开发一种廉价的三维倾斜器,并与其他微重力模拟器使用海洋分枝杆菌的比较

Joseph L. Clary, Creighton S. France, Kara R. Lind, Runhua Shi, J. Alexander, J. Richards, R. Scott, Jian Wang, Xiao-Hong Lu, Lynn Harrison
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引用次数: 1

摘要

2D和3D回转器用于模拟地球上的微重力。这些机器不断改变样品的方向,因此加速度矢量的变化速度比被监测的生物终点快。两款商用微重力模拟器分别是旋转细胞培养系统(synconon Inc.)和RPM 2.0 (Yuri),前者是一款2D摇摆器,后者是一款3D摇摆器,可以作为随机定位机或恒帧速度模式运行。我们已经开发了一种廉价的3D恒温器,可以3D打印和组装容易。为了确定模拟微重力的内帧(I)和外帧(O)速度的最佳组合,考虑了两个因素:时间平均量级和加速度矢量的分布。建立了一个计算机模型来预测帧速度在0.125转/分(rpm)和4转/分(rpm)之间组合的加速度矢量,并预测I = 1.5转/分和O = 3.875转/分的组合可以产生最佳的微重力模拟。在进一步的测试中还使用了另外两种帧速度组合:I = 0.75 rpm和O = 3.625 rpm, I = 2 rpm和O = 1.125 rpm。通过在这三种速度组合下以等速模式运行RPM 2.0, RPM 2.0算法数据证实了这些操作条件是模拟微重力的。选择海洋分枝杆菌进行生物学比较实验,因为这种细菌可以作为生物膜或浮游培养物生长。生物膜实验表明,RPM 2.0和I = 1.5 RPM和O = 3.825 RPM的3D回转器在附着的生物膜上产生了相似的结构,悬浮细菌的转录组变化与正常重力下的转录组相似。与正常重力相比,在I = 2 rpm和O = 1.125 rpm的速度下操作3D摇床,在模拟微重力方向下以25 rpm的速度操作synconon 2D摇床,可以减少浮游生物的生长,提高利福平的存活率。本研究验证了价格低廉的3D回转器,并证明了用生物实验测试实验室开发的回转器工作条件的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of an inexpensive 3D clinostat and comparison with other microgravity simulators using Mycobacterium marinum
2D and 3D Clinostats are used to simulate microgravity on Earth. These machines continuously alter the sample’s orientation, so the acceleration vector changes faster than the biological endpoint being monitored. Two commercially available microgravity simulators are the Rotary Cell Culture System (Synthecon Inc.), which is a 2D clinostat, and the RPM 2.0 (Yuri), which is a 3D clinostat that can operate as a random positioning machine or in constant frame velocity mode. We have developed an inexpensive 3D clinostat that can be 3D printed and assembled easily. To determine the optimal combination of inner (I) and outer (O) frame velocities to simulate microgravity, two factors were considered: the time-averaged magnitude and the distribution of the acceleration vector. A computer model was developed to predict the acceleration vector for combinations of frame velocities between 0.125 revolutions per minute (rpm) and 4 rpm, and a combination of I = 1.5 rpm and O = 3.875 rpm was predicted to produce the best microgravity simulation. Two other frame velocity combinations were also used in further tests: I = 0.75 rpm and O = 3.625 rpm, and I = 2 rpm and O = 1.125 rpm. By operating the RPM 2.0 in constant velocity mode at these three velocity combinations, the RPM 2.0 algorithm data confirmed that these operating conditions simulated microgravity. Mycobacterium marinum was selected for biological comparison experiments as this bacterium can grow as a biofilm or a planktonic culture. Biofilm experiments revealed that the RPM 2.0 and the 3D clinostat with I = 1.5 rpm and O = 3.825 rpm produced similar structures in attached biofilm, and similar changes in transcriptome for the bacteria in suspension compared to the normal gravity transcriptome. Operating the 3D clinostat at I = 2 rpm and O = 1.125 rpm, and the Synthecon 2D clinostat in simulated microgravity orientation at 25 rpm resulted in the same decreased planktonic growth and increased rifampicin survival compared to normal gravity. This study validates the inexpensive 3D clinostat and demonstrates the importance of testing the operating conditions of lab-developed clinostats with biological experiments.
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